9-Oxoxanthene-N,N{40 -bis(substituted)-2,7-disulfonamides

ABSTRACT

WHERE R1 is hydrogen or lower alkyl; R2 is lower alkoxy(lower)alkyl, di(lower)alkylamino (lower)alkyl, halobenzyl or morpholino(lower)alkyl; and R1 and R2 taken together with the nitrogen of the sulfonamide group are piperidino, piperazino, phenylpiperazino and morpholino. The compounds have activity as central nervous system depressants.   The disclosure is directed to 9-oxoxanthene-N,N&#39;&#39;bis(substituted)-2,7-disulfonamides which have the formula

United States Patent [191 Santilli et a1.

[ 9-OXOXANTHENE-N,N-

BIS(SUBSTITUTED)-2,7-DISULFONAMIDES [75] Inventors: Arthur A. Santilli, Havertown;

Anthony C. Scotese, King of Prussia; Stanley C. Bell, Penn Valley, all of Pa.

[73] Assignee: American Home Products Corporation, New York. NY.

[22] Filed: Feb. 12, 1973 [21] Appl. No.: 331,392

[52] US. Cl... 260/246 B; 260/268 TR; 260/293.58; 260/335; 424/248; 424/250; 424/267; 424/283 [51] Int. Cl.... C07d 7/44; C07d 29/34; C07d 87/46 [58] Field of Search 260/247.1, 246 B, 335. 260/293.58, 268 PH, 268 TR [56] References Cited UNlTED STATES PATENTS 3,067,209 12/1962 Doebel et a1, 260/247.1 P 3,073,847 1/1963 Doebel et :11. 260/2471 P 3,310,553 3/1967 Bloom et 21].... 26()/247.l P

3.75531) 8/1973 Bays 1 1 2601247.] S 3,801,598 4/1974 Pfister et a1 260/335 Pf/HZGI') Examiner-Lorraine A. Weinberger Assistant E.\'aminer--Michael Shippen Attorney, Agent, or Firm-Joseph Martin Weigman [451 Nov. 11, 1975 [57] ABSTRACT The disclosure is directed to 9-0xoxanthene-N.N'- bis(substituted)-2.7-disulfonamides which have the formula 8 Claims, N0 Drawings 1 9-OXOXANTHENE-N,N-BlS(SUBSTITUTED)-2,7- DISULFONAMIDES This invention relates to new and useful 9-oxoxan-' thene'N,N-bis(substituted)-2,7-disulfonamides and their pharmaceutically acceptable addition salts. The

chemical structure of the compounds of the invention may be schematically represented by l NO [I 2 SO h l 2 Where R is hydrogen or lower alkyl;

R is lower alkoxy (lower) alkyl, di(lower)alkylamino 25 1 ClO s H 0 c1 tto 2 ria R 0 where R and R are as defined above.

The closest known prior art is French Pat. No.

839,698 issued Apr. 1939 cs. 33 8025); Belgium Pat.

No. 759,292 issued May, 1971 (Derwent 381575); Netherlands Pat. No. 7,008,636 issued Dec. 1970 (Derwent 009885) and U.S. Pat. No. 3,532,711 issued Oct. 1970. None of the prior art cited teach or suggest the compounds of the present invention or the utility found for them.

Compounds of the present invention may be prepared by the following procedure. 9-Oxoxanthene is added to chlorosulfonic acid and heated, preferably on a steam bath, for one to five hours, preferably three hours. The reaction mixture is brought to a boil, preferably for about 5 minutes, cooled and cautiously poured on ice. A precipitate deposit may be collected and the 2 product recovered, for instance by recrystallization from benzene. The product is 2,7-di(chlorosulfonyl) xanthen-9-one.

To a solution of pyridine and an amine of Structure III is added 2,7-di(chlorosulfonyl)xanthen-9-one. The reaction mixture is heated, if necessary to produce a clear solution, and is allowed to stand at room temperature for V2 to 16 hours and then poured into water. The product separates as a solid or oily residue and may be recovered by well known means for instance by filtration, and recrystallization from a suitable organic solvent.

In the pharmacological evaluation of the biological activity of the compounds of this invention, the in vivo effects are tested as follows. The compound is administered orally or intraperitoneally to three mice (14 to 24 grams) at each of the following doses: 400, 127, 40 and 12.7 milligrams per kilogram of host body weight (MPK). The animals are watched for a minimum of 2 hours during which time signs of general stimulation, (i.e., increased spontaneous motor activity, hyperactivity on tactile stimulation, twitching), general depression (i.e., decreased spontaneous motor activity, decreased respiration) autonomic activity (i.e., miosis, mydriasis, diarrhea) are noted.

In the above test procedure the compounds of the present invention display depressant activity at 12.7 to 40.0 milligrams per kilogram of host body weight.

All of the compounds have activity as central nervous system depressants. Some have activity as interferon inducers particularly disulfonyl) bis dipiperidine. N,N'-bis(p-chlorobenzyl)- N,N-dimethyl-9-oxoxanthene-2,7-disulfonamide has activity as an anti-inflammatory and immunosuppressive agent. 4,4-(xoxanthene-2,7-diyldisulfonyl) dimorpholine has activity as an anti-inflammatory agent. N,N'-Bis(2-morpholinoethyl)9-oxoxanthene- 2,7-disulfonamide has anti-tremorine activity and trichomonacidal activity.

When the compounds of this invention are employed as described above, they may be administered alone or in combination with pharmacologically acceptable carriers, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration and standard pharmacological practice. For example, they may be administered orally in the form of tablets or capsules containing such excipients as starch, milk, sugar, certain types of clay and so forth. They may be administered sublingually in the form of troches or lozenges in which the active ingredient is mixed with sugar and corn syrups; and then dehydrated sufficiently to make it suitable for pressing into a solid form. They may be administered orally in the form of solutions or they may be injected parenterally, that is, intramuscularly, intravenously or subcutaneously. For parenteral administration they may be used 4 EXAMPLE 2 O NO ll 2!.

in the form of a sterile solution containing other solutes, for example, enough saline or glucose to make the solution isotonic.

The dose of the present pharmacological agents will vary with the form of administration and the particular compound chosen. Furthermore, it will vary with the particular subject under treatment. Generally, treatment'is initiated-with small dosages substantially less than the optimum dose of the compound. Thereafter, thedosage is increased by small increments until the optimum effect under the circumstances is reached. It will generally be found that when the composition is administered orally, larger quantities of the active agent will be required to produce the same effect as a smaller quantity given parenterally. In general, the compounds of this invention are most desirably administered at a concentration level that will generally afford effective results without cuasing any harmful or deleterious side effects.

In .order to more clearly disclose the nature of the present invention, specific examples of the practice of theinvention are hereinafter given. It should be understood, however, that this is done solely by way of example and is intended neither to delineate the scope of the invention nor to limit the ambit of the appended claims. In the examples all temperatures are given in degrees centigrade and the following abbreviations are used: g" for grams, ml for milliliters, and hr. for hours.

EXAMPLE 1 The following illustrates the preparation of a starting material 2,7-di(chlorosulfonyl)xanthen-Q-one. Ten grams of 9-oxoxanthene was added to 150 ml of chlorosulfonic acid and heated on a steam bath for 3 hr. The reaction mixture was then brought to a boil for about 5 minutes, cooled and cautiously poured on to ice. A precipitate was deposited which was collected and recrystallized twice from benzene. The product amounted to g and had amelting point of 212215.

Anal. Calcd for C H C1 O S I C, 39.71; H, 1.54; Cl, 18.03; S, 16.31. Found: C, 39.74; H, 1.70; C1, 18.44;

S, 16.53. t I

The following illustrates the preparation of 1,1 '-(9 oxoxanthene-Z,7-diyldisulfonyl)bis dipiperidinc.

To 40 ml of pyridine was added 3.4 g of piperidine and 3.9 g of 2,7-di(chlorosulfonyl)xanthen-9-one. The reaction mixture was heated for a few minutes on a hot plate until a clear solution was obtained and was then allowed to remain at room temperature for 30 minutes. The reaction mixture was poured into 700 ml of water and the yellow precipitate that was deposited was collected on a filter and washed with ethanol. There was obtained 3.5 g of product. Recrystallization from dimethylformamide gave 2.0 g of pure product having a melting point of 254-256.

The molecular formula C H N O S was assumed for the product, and based on that formula it was calculated that the elemental analysis by weight would be 56.31 percent carbon, 5.34 percent hydrogen, and 5.71 percent nitrogen. The assumed formula was determined to be accurate when it was found by analysis that the product actually contained 56.26 percent carbon, 5.38 percent hydrogen and 5.67 percent nitrogen. This may be expressed:

Anal. Calcd for C H N O S C, 56.31; H, 5.34; N, 5.71. Found: C, 56.26; H, 5.38; N, 5.67.

When evaluated in the foregoing pharmacological procedure the compound was found to be a central nervous system depressant in that it caused decreased motor activity and decreased respiration at a dose of 12.7 MPK administered parenterally.

The product was evaluated in the following procedure and found to have interferon inducing activity.

The activity of the compounds as interferon inducers was determined as follows. A dose of 2 milligrams per mouse of test compound was injected intraperitoneally. The mouse sera were harvested at 2,6, and 24 hours post-injection. The sera were assayed for interferon in cultures of L-cells (a strain of mouse fibroblasts). The interferon titer ranged from less than to equal to the interferon titer induced by 2,7-bis( 2-diethylaminoethyloxy)fluoren-9-one, dihydrochloride (Tilorone).

EXAMPLE 3 i O SO -N Q This illustrates the preparation of 4,4-(9-oxoxanthene-2,7-diyldisulfonyl)dimorpholine.

To 45 ml of pyridine was added 3.48 g of morpholine and 3.9 g of 2,7-di(chlorosulfonyl)xanthen-9-one. The

6 reaction mixture was allowed to sit for 30 minutes at This illustrates the preparation of 1,1 -(9-oxoxanroom temperature after the initial ,heat of reaction had thene-ZJ-diyIdiS uIf nyI)bis(4-phenylpiperazine). subsided. it was then poured into 800 ml of water. The T 45 m1 of p ridine was added 6.5 g of l-phenylpipproduct which precipitated from the solution was colerazine and 39 g f 2 7-di( h] 1f n[h lected on a filter and recrystallized from dimethylform- 9-one. After allowing the reaction mixture to stand at amide giving 3.4 g of product, having a melting point of room temperature for 1 hr. it was poured into 400 ml of 3083l0. water. A yellow precipitate was formed which was re- Anal. calcd for C l-l N O S C, 51.00; H, 4.48; moved by filtration and recrystallized from aqueous di- Foundi methylformamide. There was obtained 2.5 g of prod- 12.59. uct, having a melting point of 312-3l5.

When evaluated in the foregoing pharmacological Anal. Calcd for C H N O S C, 61.47; H, 5.00; N, procedure the compound was found to be acentral ner- 8.69. Found: C, 61.35; H, 5.24; N, 8.41. vous system depressant at a dose of 12.7 MPK adminis- When evaluated in the foregoing pharmacological tered parenterally. procedure the compound was found to be a central ner- The compound was also found to have anti-inflaml5 vous system depressant in that it caused decreased resmatory activity when evaluatd in the following procepiration and decreased motor activity at a dose of 12.7 dure. MPK administered parenterally.

Anti-inflammatory activity of a compound is assessed by its ability to inhibit experimentally induced edema in EXAMPLE 5 H O MQOCH CH NO S ll SO NHCH CH OMe the hind paw of the rat. Male Sprague-Dawley rats The following illustrates the preparation of N,N- weighing 120-160 grams are used as hosts. The combis(2-methoxyethyl)-9-oxoxanthene-2,7-disulfonapound to be tested is administered orally as a solution mide. or suspension in distilled water (plus two drops Tween To 50 ml of pyridine was added 4.5 g of 2-methoxye- 80) in a volume of 10 milliliters per kilogram of host thylamine followed by the porti0n-wise addition of3.93 body weight. Each compound is given to six rats and g of 2,7-di (chlorosulfonyl)xanthen-9-one. After allowvehicle along is administered to six more rats as a coning the reaction to stand overnight at room temperatrol. Sixty minutes after drug administration, edema is 35 ture, the reaction mixture was poured into 900 ml of induced by an injection of 0.05 ml of 1 percent carrawater. A precipitate was obtained which amounted to geenin solution in saline into the subplantar tissue of 6.45 g and had a melting point of 198-200. Recrystalthe rats right hind paw. Paw volume is then immedilization from N,N-dimethylformamide gave 4.0 g of ately measured volumetrically with a plethysmograph product, having a melting point of 200-203. and is measured again 3 hours later. The mean volume Anal. Calcd for C H N O S C, 48.50; H, 4.71; N, of swelling for the control group is calculated and com- 5.95; S, 13.63. Found: C, 48.49; H, 4.89; N, 6.06; S, pared to the test groups. Compounds that inhibit swell- 13.07. ing approximately 20 percent as compared to controls When evaluated in the foregoing pharmacological are considered active. Inhibition is calculated by the procedure the compound was found to be a central nerformula; vous system depressant in that it caused decreased res- Mcan vol. swelling of control-mean vol. swelling of test Percent Inhibition X 100 Mean vol. swelling of control The compound was found to cause 23 percent inhibipiration and decreased motor activity at a dose of 12.7

tion. MPK administered parenterally.

EXAMPLE 4 EXAMPLE 6 O H O NCH CH NO S H SO NHCH CH N 7 The following illustrates the preparation of N,N- bis(2-morpholinoethyl)-9-oxoxanthene-2,7-disulfonamide, hydrochloride, hemihydrate.

To 50 ml of pyridine was added 5.2 g of N(2-amino- Diamond medium (formula furnished by American Type Culture Collection,- Rockville Md.) and 0.1 ml of calf serum. Each assay tube is inoculated with 0.1 milliliters of a 48-72 hour culture of Trichomonas vaginalis ethyl) morpholine followed by the addition of 3.93 g of 5 strain ATCC No. 13972. A control assay tube contain- 2,7-di (chlorosulfonyl)xanthen-9-one. After allowing the reaction mixture to stand for 3 hours it was poured into 1.5 liters of water. A crystalline product separated which was collected and washed with ethanol. This free ing notest material is similarly inoculated. The assay tubes are gently shaken and then incubated for 48 hours at 30C. Following incubation the tubes are gently shaken and with the aid of a Pasteur pipette a base amounted to 5.5 g, and had a m ltin i t f 10 drop is deposited on a Spiers-Levy eosinophil counting 240242. The free base was added to 150 ml of ethanol and hydrogen chloride gas was bubbled into the mixture for a few minutes. The mixture was heated on a hot plate and sufficient water was added to obtain a clear solution. The reaction solution was charcoaled and filtered. The crystalline product which deposited on cooling the filtrate amounted to 3.9 g, and decomposed at 279280.

Anal. Calcd for C H C1 N O S H O/2: C, 45.32; H, 5.32; N, 8.45; Cl, 10.70; S, 9.68. Found: C, 45.26; H, 5.31; N, 8.44; Cl, 10.24 S, 9.68.

When evaluated in the foregoing pharmacological procedure the compound was found to be a central nertered parenterally.

l-l NCH CH NO The compound was found to have anti-tremorine activity by the following procedure. A literature reference is Everett et al. Science 124:79, 1956.

Compounds at a number of dose levels are administered orally (or intraperitoneally) to groups of six mice (three males and three females) at each dose level. One hour later (or one-half hour later if the compound is administered intraperitoneally) the animals are challenged with tremorine milligrams per kilogram of host body weight (MPK) administered intraperitoneally. The animals are graded at one-half hour, 1 hour and 2 hours for degree of tremors, salivation, lacrimation and diarrhea.

Protection against these symptoms is determined by a comparison with controls run simultaneously. Congentin may be used as a standard compound.

Percent antagonism at each time interval is calculated as follows:

average score (control)-average score (test) Percent antagonism average score control The compound was found to have Trichomonacidal activity in vitro when evaluated as follows.

A 12.5 milligram portion of test material (as based on the active moiety) is added to 2.5 milliliter (ml) of 1 percent phosphate buffer at pH 6. Further two-fold dilutions are made in the same buffer. A 1 milliliter volume of each dilution is transferred to small sterile screw cap assay tubes containing 3.8 milliliters of chamber. The chamber is examined by phase contrast microscopy. The number of organisms present in one square millimeter is multiplied by 5000 in order to express the count per milliliter. The difference between 15 the number of organisms present in the control tube and in the tubes containing the test material represents the relative potency of the test material and is expressed as the percentage kill at the specific dose level. Metronidazole may be used as a suitable positive con- 20 trol test material.

The compound showed a 40 percent kill.

EXAMPLE 7 0 Et II SO NHCH CH N The following illustrates the preparation of N,N- bis( 2-diethylaminoethy1)-9-oxoxanthene-2,7-disulfonamide, dihydrochloride.

To 50 ml of dry pyridine was added 9.3 g of N,N-diethy1ethylene diamine followed by the portion wise addition of 7.86 g of 2,7-di(chlorosulfonyl)xanthen-9-one. The reaction mixture was allowed to stand at room temperature for 1 hr. and was then poured into 350 ml of water. An oily residue was deposited which crystallized to give 9.5 g of free base having a melting point of l10-1l2. This material was dissolved in 150 ml of ethanol and hydrogen chloride gas was bubbled into the solution for approximately 15 minutes. A crystalline material was deposited after a few minutes. The reaction mixture was cooled in ice and filtered. The filter cake was recrystallized from ethanol giving 7.3 g of product, having a melting point of 180182.

Anal. Calcd for C H N O S 2HCl: C, 47,99; H,

6.12; N, 8.96; CI, 11.33. Found: C, 47.72; H, 6.39; N,

When evaluated in the foregoing pharmacological procedure the compound was found to be a central nervous system depressant in that it caused decreased respiration and decreased motor activity at a dose of 40 MPK administered either orally or parenterally.

The product was evaluated in the interferon induc- 9 tion procedure described in Example 2 and found to have interferon inducing activity.

EXAMPLE 8 The following illustrates the preparation of N,N'- bis(p-chlorobenzyl)-N,N-dimethyl-9-oxoxanthene- 2,7-disulfnamide.

To 40 ml of pyridine was added 6.2 g of p-chloro-N- methylbenzylamine and 3.9 g of 2,7-di(chlorosulfonyl)xanthen-9-one. A yellow precipitate was deposited. The reaction mixture was heated on a hot plate until a clear solution was obtained and then allowed to cool to room temperature. The mixture was poured into 700 ml of water. The precipitate which was deposited was collected and amounted to 6.1 g, having a melting point of 238-24l. Recrystallization from dimethylformamide gave 5.0 g of product having a melting point of 24l-243.

Anal. Calcd for C H N O Cl S C, 55.15; H, 3.83; N, 4.43. Found C, 55.15; H, 3.93; N, 4.64.

When evaluated in the foregoing pharmacological procedure the compound was found to be a central nervous system depressant at a dose of 12.7 MPK administered parenterally.

The compound was also found to bean immunosuppressive agent when evaluated as follows.

Polyarthritis is induced in male Lewis strain rats (150-20O mgs.) by the injection of a suspension of tubercle bacilli in mineral oil in the subplantar tissue of the right hind paw. Drug therapy is either begun on the day of antigen or can be started after appearance of an established arthritic syndrome (14 days). Compounds are administered daily in the form of a fine suspension by stomach tube. Body weights, left and injected right paw volumes and occurrence of arthritic nodules on the ears, tail and front paws are determined at frequent intervals over a 14 to 21 day period. All animals are then autopsied and stress organ weights, hematololgy, histopathology and biochemical studies on blood proteins are done. Active compounds will either prevent or reverse the joint swelling and associated sequella of polyarthritis with minimal toxicity.

Compounds used as standards are Cortisone, phenylbutazone, indomethacin, aspirin, and o-mercaptopurine. Literature references are Newbould, B.B. Brit. J. Pharm. Chemoth. 21:127, 1963 and Rosenthale, M.E.

10 and Nagra, C.L. Proc. Soc. exp Bio]. and Med. :149, 1967.

The compound showed 26 percent protection against acute inflammation.

OH I 3 I What is claimed is: 1. A compound selected from those having the formula R is hydrogen or lower alkyl;

R is lower alkoxy(lower)alkyl, di(lower)alkylamino (lower)alkyl, halobenzyl or morpholino(lower)alkyl; and

R and R taken together with the nitrogen of the sulfonamide group are piperidino, piperazino, phenylpiperazino and morpholino;

and their pharmaceutically acceptable acid-addition salts.

2. A compound as defined in claim 1 which is 1,] '-(9- oxoxanthene-Z,7-diyldisulfonyl)dipiperidine.

3. A compound as defined in claim 1 which is 4.4'-(9- oxoxanthene-2.7-diyldisulfonyl)dimorpholine.

4. A compound as defined in claim 1 which is 1,! '-(9- oxoxanthene-Z,7-diyldisulfonyl)bis(4-phenylpiperazine).

5. A compound as defined in claim 1 which is N,N'- bis (2-methoxyethyl)-9-oxoxanthene-2,7-disulfonamide.

6. A compound as defined in claim 1 which is N,N- bis (2-morpholinoethyl)-9-oxoxanthene-2,7-disulfonamide, dihydrochloride. hemihydrate.

7. A compound as defined in claim 1 which is N,N'- bis (Z-diethylaminoethyl)-9-oxoxanthene-2,7-disulfonamide, dihydrochloride.

8. A compound as defined in claim 1 which is N,N- bis-chlorobenzyl)-N,N-dimethyl-9-oxoxanthene-2,7-

disulfonamide.

UNITED STATES PATENT AND TRADEMARK OFFICE CERTIFICATE OF CORRECTION PATENT NO. I 3,919,211 DATED November 11, 1975 INVENTOR(S) 1 Arthur A. Santilli, Anthony C. Scotese and I, Stanley C. Bell it rs certified that error appears m the above-rdentrtred patent and that sard Letters Patent are hereby corrected as shown below:

Claim 8 should read A compound as defined in Claim 1 which is ,'bis (g-chlorobenzyl)lj,l}I '-dimethyl9 O oxoxanthene-2 7-disulfonamide Signed and gcaled this 6 [SEAL] ninth y f March 1976 Attest:

RUTH C. MASON I C. MARSHALL DANN Anestmg 0/116" ("""I'II'XSIIHIW vfPaIenIs and Trademarks 

1. A COMPOUND SELECTED FROM THOSE HAVING THE FORMULA
 2. A compound as defined in claim 1 which is 1,1''-(9-oxoxanthene-2,7-diyldisulfonyl)dipiperidine.
 3. A compound as defined in claim 1 which is 4.4''-(9-oxoxanthene-2,7-diyldisulfonyl)dimorpholine.
 4. A compound as defined in claim 1 which is 1,1''-(9-oxoxanthene-2,7-diyldisulfonyl)bis(4-phenylpiperazine).
 5. A compound as defined in claim 1 which is N,N''-bis (2-methoxyethyl)-9-oxoxanthene-2,7-disulfonamide.
 6. A compound as defined in claim 1 which is N,N''-bis (2-morpholinoethyl)-9-oxoxanthene-2,7-disulfonamide, dihydrochloride, hemihydrate.
 7. A compound as defined in claim 1 which is N,N''-bis (2-diethylaminoethyl)-9-oxoxanthene-2,7-disulfonamide, dihydrochloride.
 8. A compound as defined in claim 1 which is N,N''-bis-chlorobenzyl)-N,N''-dimethyl-9-oxoxanthene-2,7-disulfonamide. 